New Near-infrared Fluorescence Research Articles

Near‐Infrared Fluorescence of Novel Pyridinium−Cyclic Enolate Betaine Dyes π‐Expanded by Condensed Cyclic Thiophene Spacers

AbstractHerein, we report the synthesis of new near‐infrared (NIR) fluorescent dyes based on the pyridinium−cyclic enolate betaine (PCB) skeleton. The formation of the electron‐donor−π−electron‐acceptor (D−π−A) type dye with the electron‐donating N,N‐diphenyl amino group and the electron‐accepting PCB skeleton through a series of π−spacers allowed us to achieve NIR fluorescence. Particularly, a dye with a thienylisothianaphthene spacer showed NIR fluorescence with a significant intensity (λPL=822 nm, ΦPL=0.19 in DMSO). Detailed experimental and theoretical analyses suggested that the structural relaxation to a planar quinoidal form in the S1 state would be critical for intense NIR fluorescence.

Extended π-conjugated systems by external ligand-assisted C−H olefination of heterocycles: Facile access to single-molecular white-light-emitting and NIR fluorescence materials

The design and synthesis of a novel π-conjugated fluorescent framework by external ligand-assisted C−H olefination of heterocycles with excellent regioselectivity and broad substrate scope are reported herein. These novel fluorescent materials could present full-color-tunable emissions with large Stokes shifts. Furthermore, the protocol provides an opportunity to rapidly screen novel organic single-molecule white-light materials with high fluorescence quantum yields. The robust organic and low-cost white light-emitting diodes could rapidly be fabricated using the white-light-emitting material. Experimental data and theoretical calculations indicate that in the white-light dual emission the relatively short wavelength from high-lying singlet state emission and the relatively long wavelength from low-lying singlet state emission. The anti-Kasha dual-emission systems will provide a foundation for the development and application of organic single-molecule white light materials, effectively promoting the development and innovation of luminescent materials. In addition, this method demonstrated its potential application in the synthesis of new near-infrared (NIR) fluorescence materials with large Stokes shifts based on the olefination of heterocycles.

A near-infrared fluorescence probe with large Stokes shift for selectively monitoring nitroreductase in living cells and mouse tumor models

Hypoxia is commonly regarded as a typical feature of solid tumors, which originates from the insufficient supply of oxygen. Herein, the development of an efficient method for assessing hypoxia levels in tumors is strongly desirable. Nitroreductase (NTR) is an overexpressed reductase in the solid tumors, has been served as a potential biomarker to evaluate the degrees of hypoxia. In this work, we elaborately synthesized a new near-infrared (NIR) fluorescence probe (MR) to monitor NTR activity for assessment of hypoxia levels in living cells and in tumors. Upon exposure of NTR, the nitro-unit of MR could be selectively reduced to amino-moiety with the help of nicotinamide adenine dinucleotide. Moreover, the obtained fluorophore emitted a prominent NIR fluorescence, because it possessed a classical “push-pull” structure. The MR displayed several distinguished characters toward NTR, including intense NIR fluorescent signals, large Stokes shift, high selectivity and low limit of detection (46 ng/mL). Furthermore, cellular confocal fluorescence imaging results validated that the MR had potential of detecting NTR levels in hypoxic cells. Significantly, using the MR, the elevated of NTR levels were successfully visualized in the tumor-bearing mouse models. Therefore, this detecting platform based on this probe may be tactfully constructed for monitoring the variations of NTR and estimating the degrees of hypoxia in tumors.

Enhancing the Selectivity of Leucine Aminopeptidase Near-Infrared Fluorescent Probes for Assisting in Surgical Tumor Resection

Selective fluorescence imaging of analytes is a challenge for monitoring diseases as homologues interfere with the imaging agents. Leucine aminopeptidase (LAP), a kind of protease, is related to tumor pathogenesis. The known LAP fluorescent probes based on leucine recognition have limited selectivity. Herein, a selective t-butyl-alanine recognition unit for LAP through the ligand regulation strategy is prepared as a new near-infrared (NIR) fluorescent probe (DCM-LAP) having a large Stokes shift of 214 nm and a high sensitivity with a detection limit of 168 mU/L. DCM-LAP has an enhanced response toward LAP with NIR fluorescence at 656 nm based on intramolecular charge transfer. The probe is selective without being interfered with by biological enzymes including the aminopeptidase N (APN). DCM-LAP can image LAP activity in living cells. It can also visualize the cell invasion and migration processes. DCM-LAP is employed in the real-time imaging of LAP in tumor-bearing nude mice and guides in the accurate resection of breast tumors. It also distinguishes tumor tissues from normal with a high tumor-to-normal ratio (9.8). The DCM-LAP probe can thus assist in the investigations of LAP-associated clinical disease.

A multifunctional near-infrared fluorescent probe for in vitro and in vivo imaging of γ-glutamyltranspeptidase and photodynamic cancer therapy

The excessive expression of γ-glutamyltranspeptidase (GGT) is known to be closely associated with the progression, invasion and metastasis of malignant tumors, enabling GGT to be a cancer-related biomarker for cancer diagnosis. The development of the multifunctional fluorescent probe that combines GGT measurement and photodynamic therapy (PDT) into a single platform possesses exciting prospect for concurrent tumor diagnosis and treatment. In this study, we designed a new near-infrared (NIR) fluorescent probe (CyI-Glu) with therapeutic function which could realize the integration of fluorescence endogenous GGT visualization for tumor diagnosis and NIR imaging-guided tumor photodynamic therapy. CyI-Glu manifested a significant turn-on NIR fluorescence (718 nm) in response to GGT, which made it favorable for monitoring GGT activity with high selectivity and sensitivity (0.014 U/L) in vitro and in vivo. Furthermore, this GGT-responsive probe possessed improved ROS generation capacity for tumor cell apoptosis and tumor growth inhibition under NIR light irradiation. Collectively, the multifunctional fluorescent probe CyI-Glu with complementary advantages demonstrates significant potential for the diagnosis and treatment of tumors in complex organisms.

In vivo photodynamic therapy based on Near-Infrared AIE cationic polymers

• Cationic AIE polymers synthesized from AIEgen monomers with D-A-D structures. • Quaternary ammonium groups in our AIE-polymers enhance the biocompatibility for PDT. • AIE polymers with absorption maximum near 520 nm and activation by white LED light. • AIE polymers with NIR fluorescence imaging capacity and effective 1 O 2 production. • A metal-free and high atomic-efficiency polymerization method for our AIE polymers. New near-infrared (NIR) photo-responsive polymers with the merits of highly biocompatibility, penetrability and biostability, are highly desirable for cancer photodynamic therapy. Herein, a near-infrared cationic polymers (DCPN-2) featuring aggregation-induced emission (AIE) is synthesized via a ring-opening polymerization to achieve the forceful restriction of intramolecular motion, yielding nanoparticles (NPs) with diameter of about ∼ 200 nm. DCPN-2 exhibits the bright emissions in the NIR region (>650 nm) covering most of visible-light range and effectively generates reactive oxygen species (ROS) under light irradiation. Importantly, the inhibition of tumor growth upon DCPN-2 treatment under light irradiation was demonstrated both in vitro and in vivo. Collectively, this study not only provides potential NIR AIE-polymers for cancer phototheranostics, but also presents a novel strategy of the design of biocompatible photo-responsive polymers for future biomedical applications.

Development of a NIR fluorescent probe for detection of cysteine and its application in bioimaging

In this work, A new near-infrared (NIR) fluorescent probe NIR-L based on dicyanoisophorone was rationally designed, synthesized and characterized for the detection of cysteine (Cys). This probe exhibited highly selective and sensitive response to Cys over other analytes including amino acids, cations and anions under physiological conditions, which can be identified by naked eyes. It was found that the probe showed NIR fluorescence turn-on response at 714 nm toward Cys with a large Stokes shift of 174 nm and a very low detection limit of 28.6 nM. The results of 1H NMR and DFT calculation further confirmed that the addition of Cys triggered a reaction between Cys and NIR-L, leading to the release of a NIR fluorescent dye NIR-OH. Moreover, NIR-L was successfully used to realize the in vivo Cys bioimaging in HeLa cells and mice. Significantly, the sensing system was applied in imaging of Cys in B16-F10 tumor-bearing mice.

An eco-friendly near infrared fluorescence molecularly imprinted sensor based on zeolite imidazolate framework-8 for rapid determination of trace trypsin

This report depicts an environment-friendly molecularly imprinted fluorescence sensor for rapid determination of trypsin based on new near infrared (NIR) fluorescence carbon dots and metal–organic frameworks. The novel NIR fluorescence carbon dots were facilely synthesized from green plants by one-step solvothermal treatment, and the imprinted sensor was prepared by sol–gel under mild condition. The imprinted NIR fluorescence sensor was characterized by Fourier-transform infrared spectrometry, energy disperse spectroscopy, scanning electron microscopy, X-ray diffraction and transmission electron microscopy. The results showed the introduction of zeolite imidazolate framework-8 can improve the fluorescence response sensitivity and response speed. Under optimized conditions, the fluorescence quenching of the imprinted NIR fluorescence sensor was linear relationship with the increasing trypsin concentration in the range of 0.05–9.0 ng/L with a detection limit of 8.8 pg/L. The developed imprinted fluorescence sensor can selectively recognize trypsin in 1.5 min, which was applied to rapidly and sensitively detect trypsin in serum and urine samples successfully with the recoveries of 95.3–104.1% and 97.7–105.2%, respectively. The as-prepared imprinted fluorescence sensor provides a viable and eco-friendly method for rapidly sensitive detection of trace bio-macromolecule in complex samples.

A novel near-infrared-emitting aza-boron-dipyrromethene-based remarkable fluorescent probe for Hg2+ in living cells

A new near-infrared (NIR)-emitting aza-boron-dipyrromethene dye with two electron-donating amino groups at 1- and 7-positions has been prepared via several steps of reactions. This probe showed a NIR absorption at 748 nm with an obvious shoulder peak at 634 nm in CH3CN/H2O. Interestingly, a NIR fluorescence emission at 843 nm was observed with a large Stokes shift of 95 nm. This novel NIR-emitting aza-boron-dipyrromethene dye was further investigated as a Hg2+-sensing fluorescent probe, which selectively bound to Hg2+, showing a blue-shifted and sharp absorption band at 695 nm with the disappearance of the shoulder peak at 634 nm. Correspondingly, the color change could be easily seen from blue to green. Interestingly, the emission exhibited an absolutely “turn-on” peak at 725 nm with a significant blue shift by 118 nm (from 843 to 725 nm), due to the efficient inhibition of the intramolecular-charge-transfer process arising from two amino groups. This probe was finally introduced to Hela cells, showing a “OFF-ON” NIR emission upon exposure to Hg2+. The overall results confirmed that this novel NIR-emitting aza-boron-dipyrromethene fluorescent probe with a large Stokes shift could serve as a colorimetric and fluorescent “turn-on” sensor for Hg2+ in both solutions and living cells.

In situ visualization of peroxisomal viscosity in the liver of mice with non-alcoholic fatty liver disease by near-infrared fluorescence and photoacoustic imaging.

Non-alcoholic fatty liver disease (NAFLD) can gradually develop into hepatic failure, and early diagnosis is crucial to improve treatment efficiency. The occurrence of NAFLD is closely related to lipid metabolism. Peroxisomes act as the first and main site for lipid metabolism in the hepatocytes, so abnormal lipid metabolism might directly affect peroxisomal viscosity. Herein, we developed a new near-infrared fluorescence (NIRF) and photoacoustic (PA) imaging probe (PV-1) for the real-time visualization of peroxisomal viscosity in vivo. This PV-1 encompasses the malononitrile group as the rotor, which emits strong NIRF (at 705 nm) and PA (at 680 nm) signals when rotation is hindered as viscosity increases. Through dual-mode imaging, we discovered distinctly higher viscosity in the liver of NAFLD mice for the first time. We further found the remarkable amelioration of NAFLD upon treatment with N-acetylcysteine (NAC). Therefore, we anticipate that the PV-1 imaging method is promising for the early diagnosis and prognostic evaluation of NAFLD.

Rapid detection of CO in vitro and in vivo with a ratiometric probe showing near-infrared turn-on fluorescence, large Stokes shift, and high signal-to-noise ratio

A new near-infrared (NIR) fluorescent probe was developed for the detection of CO in living systems. This probe is based on a unique cyanine fluorophore and uses an allyl ether moiety as the reaction site. Optical studies in solution show that this probe has good water solubility and possesses desirable sensing properties for CO including rapid response, high selectivity and sensitivity, large Stokes shift (123 nm), distinct colorimetric changes, and significant turn-on NIR fluorescence changes at 715 nm with high signal-to-noise ratio. Ratiometric fluorescent detection of CO with this probe at 715 and 570 nm is also applicable. The sensing mechanism was proven to be a process of recovery of the fluorophore by removing the allyl group via Tsuji-Trost reaction. Furthermore, this probe was applied for detecting CO in living cells and animals with good bioimaging performance. Moreover, indicated by this probe, more CO could be produced under oxidative stress conditions. All the results indicate that this probe can be applied as a promising new tool for in vitro and in vivo detection of CO.

A new colorimetric and near-infrared fluorescent probe for biothiols and its application to image GSH in living cells and zebrafish

A new near-infrared (NIR) fluorescent probe based on 2-Methylchromone was exploited for relay recognition of Cu2+ and biothiols. The mechanism involved in complex between metal ion and probe, and subsequent displacement reaction, which make the probe can detect GSH by NIR fluorescence turn on emission. Probe showed a fluorescence band at 718 nm in PBS-EtOH (3:2, v/v, 10 mM PBS, pH 7.4) solution. Upon the addition of Cu2+, a large fluorescence quenching was observed due to the coordination between probe 1 and Cu2+. The 1-Cu2+ could be used to detect biothiols utilizing high affinity of biothiols with Cu2+. This “on-off-on” fluorescent probe was also successfully applied to image GSH in MCF-7 cells and zebrafish.

Development of Theragnostic Tool Using NIR Fluorescence Probe Targeting Mitochondria in Glioma Cells.

Because mitochondria are essential organelles for regulating energy homeostasis and intrinsic apoptosis, the perturbation of mitochondrial functions has been considered as an anticancer treatment. In this study, a new near-infrared (NIR) fluorescent probe, SiR-Mito11 was developed as a theragnostic agent for brain tumor by targeting mitochondria. SiR-Mito11 exhibited potential anticancer activity against glioma cells but tolerance in normal neuronal cells. We further confirmed that the selective accumulation of SiR-Mito11 in glioma cells disrupted mitochondria membrane potential, followed by apoptotic cell death.

Highly Stable and NIR Luminescent Ru-LPMSN Hybrid Materials for Sensitive Detection of Cu2+ in Vivo.

Herein, new near-infrared (NIR) luminescent ruthenium complexes were prepared for detecting Cu2+ ions. Then, ruthenium complex hybrid nanomaterials (Ru-LPMSNs) were fabricated successfully by imbedding the ruthenium complex into mesoporous silica nanoparticles. Benefiting from the novel large-pore mesoporous structure and good adsorbility of LPMSNs, Ru-LPMSN hybrid materials showed a significantly enhanced fluorescence intensity and stability. NIR fluorescence of Ru-LPMSNs was rapidly quenched by Cu2+ ions. Ru-LPMSNs also showed high Cu2+ ion selectivity and sensitivity as a sensor. The detection limit of Cu2+ ions was 10 nM with a wide linear relationship between the fluorescence intensity of Ru-LPMSNs and the concentration of Cu2+ ions. The mechanism of fluorescence quenching might be that the combination of the ruthenium complex and Cu2+ ions constrained the photoinduced electron-transfer process. Furthermore, Ru-LPMSNs dramatically increased the fluorescence signals in cells and achieved Cu2+-ion detection. Ru-LPMSNs had different tissue affinities and could monitor distribution of exogenous Cu2+ ions in vivo. Moreover, Ru-LPMSNs realized direct and rapid detection of Cu2+-ion content in serum. These results indicated the potential applications of the prepared nanomaterials as Cu2+ detection agents.

Intraoperative Ureter Visualization Using a Novel Near-Infrared Fluorescent Dye.

Ureters can be accidentally severed during pelvic surgeries, significantly prolonging the times in the operating room to allow for complete repair of damaged ureters and leading to significant morbidities associated with consequent ureter obstruction and possible kidney dysfunction. In an effort to prevent these complications, light-emitting stents and urine-excreted dyes have been introduced to illuminate the ureter during surgery. However, problems with mechanical insertion, ureter spasm, image contrast, and localized injection have limited interest in their clinical applications. We report here the synthesis and characterization of a new near-infrared (NIR) fluorescent dye (UreterGlow) that can be injected systemically but is excreted primarily through the renal system, allowing ureter imaging with an NIR fluorescence camera. Following intravenous injection of 0.1 mg/kg UreterGlow, we have monitored the flow of UreterGlow through the proximal, medial, and distal segments of the ureter. The timing of ureter visualization was calculated from the time of injection of the drug. The null hypothesis was that "Visualization of the ureter in pigs is possible 60 min after administration of UreterGlow using an NIR camera". UreterGlow displayed excitation and emission maxima of λex = 800 nm and λem = 830 nm in phosphate buffered saline, pH 7.4, and could be imaged in the urinary tract in mice. Shortly after injection of UreterGlow into Yorkshire pigs, peristalsis of the ureter could be observed. The distal ureter could be visualized under NIR illumination after 60 min with constant fluorescence in all five pigs for >2 h. The same ureters could not be seen using visible light ( X2, p = 0.0001). Because both excitation and emission of UreterGlow occurs at >30 nm longer wavelength than most tumor-imaging fluorescent dyes, it should be possible to distinguish ureter fluorescence from tumor fluorescence with this dye.

Long-term prevalence of NIRF-labeled magnetic nanoparticles for the diagnostic and intraoperative imaging of inflammation

Inflammation is a very common disease worldwide. In severe cases, surgery is often the method of choice. Today, there is a general need for the implementation of image-based guidance methodologies for reliable target resection. We investigated new near infrared fluorescence (NIRF)-nanoparticles (NPs) as a simple but effective bimodal magnetic resonance imaging (MRI) and optical contrast agent for diagnosis and intraoperative imaging of inflammation. Physicochemical analysis revealed that these NPs were highly fluorescent with similar characteristics like unlabeled NPs (hydrodynamic diameter about 130 nm and zeta potential about −10 mV). NP-uptake and NIR-dye labeling was biocompatible to macrophages (no impact on cellular ATP and reactive oxygen species production). These cells could successfully be tracked with MRI and NIRF-optical imaging. I.v. injection of fluorescent NPs into mice led to highly specific T2-weighted signal of edema due to uptake by phagocytic cells and subsequent migration to the site of inflammation. NIRF signals of the edema region were well detectable for up to 4 weeks, underlining the potential of the NPs for systematic planning and flexible time scheduling in intraoperative applications. NPs were degraded over a time period of 12 weeks, which was not altered due to inflammation. Redistribution of iron might be primarily due to inflammation and not to the presence of NPs per se in a concentration suitable for imaging. Our findings highlight the potential of the NPs to be used as a suitable tool for pre- and intraoperative imaging of inflammation.

N-annulated perylene-substituted and fused porphyrin dimers with intense near-infrared one-photon and two-photon absorption.

Fusion of two N-annulated perylene (NP) units with a fused porphyrin dimer along the S0-S1 electronic transition moment axis has resulted in new near-infrared (NIR) dyes 1 a/1 b with very intense absorption (ε>1.3×10(5) M(-1) cm(-1)) beyond 1250 nm. Both compounds displayed moderate NIR fluorescence with fluorescence quantum yields of 4.4×10(-6) and 6.0×10(-6) for 1 a and 1 b, respectively. The NP-substituted porphyrin dimers 2 a/2 b have also been obtained by controlled oxidative coupling and cyclodehydrogenation, and they showed superimposed absorptions of the fused porphyrin dimer and the NP chromophore. The excited-state dynamics of all of these compounds have been studied by femtosecond transient absorption measurements, which revealed porphyrin dimer-like behaviour. These new chromophores also exhibited good nonlinear optical susceptibility with large two-photon absorption cross-sections in the NIR region due to extended π-conjugation. Time-dependent density functional theory calculations have been performed to aid our understanding of their electronic structures and absorption spectra.

In vivo detection of macrophage recruitment in hind-limb ischemia using a targeted near-infrared fluorophore.

Macrophages are an essential component of the immune system and have protective and pathogenic functions in various diseases. Imaging of macrophages in vivo could furnish new tools to advance evaluation of disease and therapies. Critical limb ischemia is a disease in which macrophages have considerable pathogenic roles, and are potential targets for cell-based immunotherapy. We sought to develop a new near-infrared fluorescence (NIRF) imaging probe to target macrophages specifically in vivo in various pathological states, including hind-limb ischemia. We rapidly screened the photostable cyanine-based NIRF library against different blood cell lines. The identified monocyte/macrophage-selective hit was tested in vitro in live-cell labeling assay. Non-invasive NIRF imaging was performed with murine models of paw inflammation by lipopolysaccharide challenge and hind-limb ischemia with femoral artery ligation. in vivo macrophage targeting was further evaluated using intravital microscopy with Csf1r-EGFP transgenic mice and immunofluorescent staining with macrophage-specific markers. We discovered MF800, a Macrophage-specific near-infrared Fluorophore, which showed selective live-cell imaging performance in a panel of cell lines and primary human blood samples. MF800 outperforms the clinically-available NIRF contrast agent ICG for in vivo specificity in paw inflammation and hind-limb ischemia models. We observed a marked overlap of MF800-labeled cells and EGFP-expressing macrophages in intravital imaging of Csf1r-EGFP transgenic mice. In the histologic analysis, MF800-positive cells also expressed the macrophage markers CD68 and CD169. NIRF imaging showcased the potential of using MF800 to understand macrophage behavior in vivo, characterize macrophage-associated diseases, and may help in assessing therapeutic responses in the clinic.

Near-infrared photochromic behavior in a donor–acceptor type diarylethene modulated by the cyanide anion

Two new near-infrared (NIR) photochromic compounds, containing both electron-donating (D) and accepting groups (A), have been developed and their NIR photochromic and fluorescence performances can be modulated by the cyanide anion. Such a colorimetric cyanide sensor has been achieved based on this donor-photochromic bridge-acceptor system.

Near-Infrared Emitting Polymer Nanogels for Efficient Sentinel Lymph Node Mapping

Sentinel lymph node (SLN) mapping has been widely used to predict the metastatic spread of primary tumor to regional lymph nodes in clinical practice. In this research, a new near-infrared (NIR)-emitting polymer nanogel (NIR-PNG) having a hydrodynamic diameter of about 30 nm, which is optimal for lymph node uptake, was developed. The NIR-emitting polymer nanoprobes were designed and synthesized by conjugating IRDye800 organic dye to biodegradable pullulan-cholesterol polymer nanogels. The NIR-PNG nanoprobes were found to be photostable compared with the IRDye800-free dye at room temperature. Upon intradermal injection of the NIR-PNG into the front paw of a mouse, the nanoprobes entered the lymphatic system and migrated to the axillary lymph node within 2 min. The NIR fluorescence signal intensity and retention time of NIR-PNG in the lymph node were superior to the corresponding properties of the IRDye800-free dye. A immunohistofluorescence study of the SLN resected under NIR imaging revealed that the NIR-PNG nanoprobes were predominantly co-localized with macrophages and dendritic cells. Intradermal injection of NIR-PNG nanoprobes into the thigh of a pig permitted real-time imaging of the lymphatic flow toward the SLN. The position of the SLN was identified within 1 min with the help of the NIR fluorescence images. Taken together, the experimental results demonstrating the enhanced photostability and retention time of the NIR-PNG provide strong evidence for the potential utility of these polymer probes in cancer surgery such as SLN mapping.